Gibson D. (2009) Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides. Nucleic Acids Res, 37:6984–6990.
When producing large, synthetic genes it is frequently necessary to join many chemically synthesized DNA elements, which is often impossible with traditional blunt- and cohesive-end cloning methods. Dr Dan Gibson, at Synthetic Genomics Inc., is a pioneer in developing methods for assembling genes from pools of synthetic, single- and double-stranded DNA. In this article, Dr Gibson describes an interesting protocol for assembling high-fidelity Ultramer® Oligonucleotides
with overlapping sequences into large constructs using Saccharomyces cerevisiae
. The method can be used with both single- (Ultramer Oligonucleotides
) and double-stranded DNA (IDT gBlocks® Gene Fragments
). In the article, Isothermal Assembly: Quick, Easy Gene Construction
, we discussed the more recently developed Gibson Assembly™ Method, which uses a unique enzyme mix to rapidly assemble large synthetic genes.